磁珠法和煮沸法核酸提取在HPV-DNA分型中的比较  

Comparison of magnetic beads method and boiling method for nucleic acid extraction in HPV-DNA genotyping

作　　者：许琼 李文静[1] 田芯瑗 徐红星[1] 吴冬生[1] 许华英[1] XU Qiong;LI Wenjing;TIAN Xinyuan;XU Hongxing;WU Dongsheng;XU Huaying(Department of Clinical Laboratory,Affiliated Suzhou Municipal Hospital,Nanjing Medical University,Suzhou,Jiangsu 215168,China;Faculty of Medical Laboratory Technology,Medical College,Suzhou University,Suzhou,Jiangsu 215000,China)

机构地区：[1]南京医科大学附属苏州市立医院本部检验科,215168 [2]苏州大学医学院,江苏苏州215000

出　　处：《检验医学与临床》2018年第6期764-767,共4页Laboratory Medicine and Clinic

基　　金：江苏省苏州市科技计划指导性项目(SYSD2016112)

摘　　要：目的比较基于Pre-NAT全自动核酸提取系统的磁珠法与基于手工的煮沸法核酸提取在人乳头瘤病毒(HPV)-DNA分型中的应用。方法收集96份宫颈脱落细胞标本用于两种DNA提取方法检测结果的比对;利用同一混合阳性标本分装后加入系列水平的血红蛋白(Hb),评价抗Hb干扰能力;收集30份临床棉拭子标本,分析两种方法对皮肤破溃或男性生殖道分泌物DNA的提取效率。结果 96份临床标本,两种方法检测结果一致率为91.7%(88/96);磁珠法内参Globin及HPV各亚型的信号值均显著高于煮沸法(t=8.807、4.676,P<0.05)。Hb水平>17g/L时煮沸法HPV-DNA分型失败,而磁珠法信号值不受Hb干扰。30份棉拭子标本,磁珠法和煮沸法检测成功率均为80.0%,阳性率分别为54.2%、41.7%(P=0.564)。结论相比手工提取的煮沸法,基于Pre-NAT全自动病毒核酸提取系统的磁珠法具有操作简便、规范、通量高等优点。两种方法具有一定可比性,磁珠法核酸提取效率高于煮沸法,且不受Hb干扰,更适于临床应用。Objective To compare the application of magnetic beads method based on Pre-NAT automatic nucleic acid extraction system and the manual boiling method in HPV-DNA genotyping.Methods Ninty-six clinical exfoliated cells samples were collected for the comparison between the two nucleic acid extraction methods;the same mixed positive sample was used after subpackage and serial levels of hemoglobin(Hb)were added for evaluating the anti-interference ability;30 clinical cotton swab specimens were collected for analyzing the DNA extraction efficiency of skin lesion or male genital tract secretions.Results Among 96 clinical samples,the concordance rate of detection results by two methods was 91.7%(88/96);the signal values of Globin and HPV subtypes in the magnetic beads method were significant higher than those in the boiling method(t=8.807,4.676,P<0.05).HPV-DNA genotyping failed in boiling method when the Hb level>17 g/L,while the signal values of magnetic beads were free from the Hb interference.The detection success rates of 30 cotton swab specimens were 80.0%,and the positive rates were 54.2%and 41.7%respectively(P=0.564).Conclusion Compared to manual extraction boiling method,the magnetic beads method based on Pre-NAT automatic nucleic extraction system has the advantages of simple operation,normalization and higher throughput.The two methods have certain comparability.Magnetic beads method has higher nucleic acid extraction rate than the manual boiling method without Hb interference and is more suitable for clinical application.

关 键 词：人乳头瘤病毒 核酸 提取 磁珠法 煮沸法 

分 类 号：R446.9[医药卫生—诊断学]